Watermelon line ca9

ABSTRACT

A watermelon cultivar, designated watermelon line CA9, is disclosed. The invention relates to the seeds of watermelon line CA9, to the plants of watermelon line CA9 and to methods for producing a watermelon plant by crossing the watermelon line CA9 with itself or another watermelon cultivar. The invention further relates to methods for producing triploid, seedless watermelon fruit using watermelon line CA9 as a parent of the pollenizer plant. This invention also relates to watermelon cultivars or breeding cultivars and plant parts derived from watermelon line CA9, to methods for producing other watermelon cultivars, lines or plant parts derived from watermelon line CA9 and to the watermelon plants, varieties, and their parts derived from the use of those methods. The invention further relates to hybrid watermelon seeds, plants, and plant parts produced by crossing watermelon line CA9 with another watermelon cultivar.

BACKGROUND OF THE INVENTION

The present invention relates to a new watermelon (Citrullus lanatus)cultivar designated watermelon line CA9. All publications cited in thisapplication are herein incorporated by reference.

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis and definitionof problems and weaknesses of the current germplasm, the establishmentof program goals, and the definition of specific breeding objectives.The next step is selection of germplasm that possess the traits to meetthe program goals. The goal is to combine in a single variety or hybridan improved combination of desirable traits from the parental germplasm.These important traits may include increased head size and weight,higher seed yield, improved color, resistance to diseases and insects,tolerance to drought and heat, and better agronomic quality.

Watermelon is a member of the Cucurbitaceae family and is a vine-likeflowering plant thought to have originated in southern Africa.Watermelon is an annual plant with long, weak, trailing or climbingstems that is grown for its large edible fruit. The fruit has a thickrind and fleshy center that is red, orange, pink, yellow, green orwhite. The fruit is rich in vitamins A and C and can be eaten raw orcooked in various ways. There are over 1,200 varieties of watermelonworldwide, which range in weight from less than one to more than 90kilograms.

Successful watermelon production depends on attention to variouscultural practices. This involves soil management practices with specialattention to proper fertilization, crop establishment with appropriatespacing, weed control, the introduction of bees for pollination, andsuitable pollenizers for seedless watermelon, irrigation and pestmanagement. Watermelon fruit size and shape; rind color; thickness andtoughness; seed size, color and number; and flesh color, texture,soluble solids and freedom from fruit defects are all importantcharacteristics to be considered in selection of watermelon varieties.In addition, seedless watermelons should be free of hard seeds and haveundeveloped seeds that are small and innocuous.

Watermelon pollination is essential to the production of fruit. Theflowers of watermelon plants are unisexual, with male and female flowersoccurring on the same plant (monoecious). In order to set fruit, pollenfrom the male flower must be transferred to a female flower on thatplant or another plant in the field. This pollen transfer isaccomplished by several naturally occurring insects, but mosteffectively by the honeybee.

Seedless watermelon plants are triploid and are produced by crossing atetraploid (2n=4x=44 chromosomes) inbred line as the female parent witha diploid (2n=2x=22) inbred line as the male parent of the hybrid; theresulting hybrid is a triploid (2n=3x=33). Triploid plants have threesets of chromosomes, and three sets cannot be divided evenly duringmeiosis. This results in nonfunctional female and male gametes althoughthe flowers appear normal. Since the triploid hybrid is female sterile,the fruit induced by pollination tend to be seedless. As the pollen intriploid male flowers is not viable and female flowers in triploidplants require viable pollen to set fruit, it follows that there must beseparate diploid (seeded) pollenizer plants available to provide pollen.

Triploid watermelon is mainly pollinated by bees and other insects thathop from flower to flower and distribute pollen from seeded pollenizerplants to triploid hybrid plants. Because watermelon flowers open onlyfor a short time, it is essential that bees and pollens are presentduring pollination. It is also essential that the full-flowering periodof the seedless plants (which takes about 3-4 weeks) should match withthe full-flowering period of the pollenizer plants, in order to haveplenty of pollen available during pollination. Lack of pollen duringfull flowering of seedless watermelon plants will have negative effecton the total yield and fruit quality. Therefore, early-maturing seedlesswatermelon hybrids should be combined with early and prolonged-floweringpollenizer plants to achieve high yield and quality watermelonproduction in commercial production fields.

Seeded watermelon plants take up space, nutrients and water in the fieldthat farmers would rather devote to seedless plants. As a results,farmers have increasingly turned to varieties of seeded watermelons thatproduce pollen to fertilize the seedless plants, but that also grow verysmall, inedible fruit that does not need to be harvested and does nottake up much space in the field. These varieties are called“pollenizers” because they are grown solely to provide pollen for theseedless watermelons.

Therefore, developing improved inbred watermelon lines having anincreased number of male flowers and an increased length of maleflowering period, and producing an increased yield of marketabletriploid fruit when used as a parent of a pollenizer, is highlydesirable.

The foregoing examples of the related art and limitations relatedtherewith are intended to be illustrative and not exclusive. Otherlimitations of the related art will become apparent to those of skill inthe art upon a reading of the specification.

SUMMARY OF THE INVENTION

The following embodiments and aspects thereof are described inconjunction with systems, tools, and methods which are meant to beexemplary and illustrative, not limiting in scope. In variousembodiments, one or more of the above-described problems have beenreduced or eliminated, while other embodiments are directed to otherimprovements.

According to the invention, there is provided a novel inbred watermeloncultivar designated watermelon line CA9. This invention thus relates tothe seeds of watermelon line CA9, to the plants of watermelon line CA9,and to methods for producing a watermelon plant produced by crossing thewatermelon line CA9 with itself or another watermelon plant, to methodsfor producing a watermelon plant containing in its genetic material oneor more transgenes, and to the transgenic watermelon plants produced bythat method. This invention also relates to methods for producing otherwatermelon cultivars derived from watermelon line CA9 and to thewatermelon cultivar derived by the use of those methods. This inventionfurther relates to hybrid watermelon seeds and plants produced bycrossing watermelon line CA9 with another watermelon variety, whereinwatermelon line CA9 is used as the male and/or the female parent.

In another aspect, the present invention provides regenerable cells foruse in tissue culture of watermelon line CA9. The tissue culture willpreferably be capable of regenerating plants having essentially all ofthe physiological and morphological characteristics of the foregoingwatermelon plant, and of regenerating plants having substantially thesame genotype as the foregoing watermelon plant. Preferably, theregenerable cells in such tissue cultures will be callus, protoplasts,meristematic cells, cotyledons, hypocotyl, leaves, pollen, embryos,roots, root tips, anthers, pistils, shoots, stems, petiole flowers, andseeds. Still further, the present invention provides watermelon plantsregenerated from the tissue cultures of the invention.

Another aspect of the invention is to provide methods for producingother watermelon plants derived from watermelon line CA9. Watermelonplants derived by the use of those methods are also part of theinvention.

In another aspect, the present invention provides methods for producingtriploid, seedless watermelon fruit when watermelon line CA9 is used asa parent of a diploid watermelon pollenizer plant. The method comprisesplanting a field with triploid watermelon plants and/or seeds; obtainingdiploid pollenizer watermelon plants and/or seeds for pollinizing thetriploid watermelon plants and/or seeds, wherein at least one parent ofthe diploid pollenizer plant is watermelon line CA9; planting thepollenizer watermelon plants and/or seeds in the field of triploidwatermelon plants and/or seeds; allowing pollination of the triploidwatermelon plants by pollen of the pollenizer watermelon plants toobtain triploid, seedless watermelon fruit; and harvesting the triploid,seedless watermelon fruit. The planting and seedlings ratios of diploidto triploid plants were approximately equal to or less than 1 diploidpollenizer watermelon plant to 3, 4, 5, or 6 triploid watermelon plants.

The invention also relates to methods for producing a watermelon plantcontaining in its genetic material one or more transgenes and to thetransgenic watermelon plant produced by those methods.

In another aspect, the present invention provides for single geneconverted plants of watermelon line CA9. The single transferred gene maypreferably be a dominant or recessive allele. Preferably, the singletransferred gene will confer such traits as male sterility, herbicideresistance, insect or pest resistance, modified fatty acid metabolism,modified carbohydrate metabolism, resistance for bacterial, fungal, orviral disease, male fertility, enhanced nutritional quality, andindustrial usage. The single gene may be a naturally occurringwatermelon gene or a transgene introduced through genetic engineeringtechniques.

The invention further provides methods for developing watermelon plantsin a watermelon plant breeding program using plant breeding techniquesincluding recurrent selection, backcrossing, pedigree breeding,restriction fragment length polymorphism enhanced selection, geneticmarker enhanced selection, and transformation. Seeds, watermelon plants,and parts thereof, produced by such breeding methods are also part ofthe invention.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by reference bystudy of the following descriptions.

DETAILED DESCRIPTION OF THE INVENTION

In the description and tables which follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

Allele. The “allele” is any of one or more alternative forms of a gene,all of which relate to one trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy corresponding loci on apair of homologous chromosomes.

Backcrossing. “Backcrossing” is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents, for example, a firstgeneration hybrid F₁ with one of the parental genotype of the F₁ hybrid.

Cotyledon. One of the first leaves of the embryo of a seed plant;typically one or more in monocotyledons, two in dicotyledons, and two ormore in gymnosperms.

Cross-pollination. Fertilization by the union of two gametes fromdifferent plants.

Diploid. A cell or organism having two sets of chromosomes.

Essentially all the physiological and morphological characteristics. Aplant having essentially all the physiological and morphologicalcharacteristics means a plant having the physiological and morphologicalcharacteristics of the recurrent parent, except for the characteristicsderived from the converted gene.

Explosive rind. A trait (e) that causes the fruit rind of watermelon toburst or split when cut. Used to make fruit easily crushed by harvestcrews for pollinator cultivars that have small fruit not intended forharvest.

Gene. As used herein, “gene” refers to a segment of nucleic acid. A genecan be introduced into a genome of a species, whether from a differentspecies or from the same species, using transformation or variousbreeding methods.

Gene converted (conversion). “Gene converted” or “Single gene converted”(or conversion) plant refers to plants which are developed by a plantbreeding technique called backcrossing wherein essentially all of thedesired morphological and physiological characteristics of an inbred arerecovered in addition to the one or more genes transferred into theinbred via the backcrossing technique or via genetic engineering.

Haploid. A cell or organism having one set of the two sets ofchromosomes in a diploid.

Linkage. Refers to a phenomenon wherein alleles on the same chromosometend to segregate together more often than expected by chance if theirtransmission was independent.

Locus. A defined segment of DNA.

Pedigree breeding/selection. “Pedigree breeding” is a breeding methodused during the inbreeding of populations of self- and cross-pollinatedspecies for the development of desirable homogeneous lines. Pedigreeselection generally begins with an F₂ population and continues untilhomogeneous lines are developed.

Petiole. “Petiole” means the stalk of a leaf, attaching the leaf bladeto the stem.

Plant. “Plant” includes plant cells, plant protoplasts, plant ovules,plant cells of tissue culture from which C. lanatus plants can beregenerated, plant calli, plant clumps and plant cells that are intactin plants, or parts of plants such as pollen, flowers, seeds, leaves,stems, fruit, rind, flesh and the like.

Pollenizer. Refers to diploid, seeded watermelon that produce pollenused to fertilize triploid, seedless watermelon plants and also growvery small, inedible fruit that does not need to be harvested.

Quantitative Trait Loci. “Quantitative Trait Loci” (QTL) refers togenetic loci that control to some degree, numerically representabletraits that are usually continuously distributed.

Regeneration. “Regeneration” refers to the development of a plant fromtissue culture.

RHS. “RHS” refers to the Royal Horticultural Society of England whichpublishes an official botanical color chart quantitatively identifyingcolors according to a defined numbering system. The chart may bepurchased from Royal Horticulture Society Enterprise Ltd., RHS Garden;Wisley, Woking; Surrey GU236QB, UK.

Scion. Refers to a detached plant shoot containing buds, flowers orfruits that is used for grafting to stock or rootstock. The scioncontains the desired genes to be duplicated in future production by thestock/scion plant.

Single gene converted. “Single gene converted” or “conversion plant”refers to plants which are developed by a plant breeding techniquecalled backcrossing or via genetic engineering wherein essentially allof the desired morphological and physiological characteristics of a lineare recovered in addition to the single gene transferred into the linevia the backcrossing technique or via genetic engineering.

Stem. “Stem” means the above ground structures that have vascular tissueand that support, for example, leaves, flowers, seed, fruit, etc. Thestem is normally divided into nodes and internodes, the nodes hold budswhich grow into for example, one or more leaves, inflorescence(flowers), cones or other stems (or branches), while the internodes actas spaces that distance one node from another.

Tetraploid. A cell or organism having four sets of chromosomes.

Triploid. A cell or organism having three sets of chromosomes.

Yield. “Yield” means the total weight in kilograms of marketableharvested fruit from an experimental plot or field.

Watermelon line CA9 is an inbred diploid watermelon line that producesan increased number of male flowers when compared to commercial andexperimental diploid watermelon lines. When used as a male parent toproduce diploid watermelon pollenizer plants, watermelon line CA9produces pollenizer plants having 1) an increased number of male flowersand 2) an increased length of male flowering period when compared to theclosest commercial diploid varieties, 3) an increased yield ofmarketable triploid fruit when used as a pollenizer and 4) a distinctiverind pattern. Watermelon line CA9 may be a male and/or female parentwhen used for the production of hybrid watermelon plants. Additionally,watermelon line CA9 has the explosive rind trait.

Watermelon line CA9 has shown uniformity and stability for the traits,within the limits of environmental influence for the traits. It has beenself-pollinated a sufficient number of generations with carefulattention to uniformity of plant type. The line has been increased withcontinued observation for uniformity. No variant traits have beenobserved or are expected in watermelon line CA9.

Watermelon line CA9 has the following morphological and physiologicalcharacteristics (based primarily on data collected in Woodland, Calif.).

TABLE 1 VARIETY DESCRIPTION INFORMATION Plant:   Species: Citrulluslanatus var. lanatus   Adaptation: Most U.S. areas   Relative maturity:70 days   Ploidy: Diploid Stem:   Number of main stems at crown: 4,including stem holding fruit   Shape: Round   Diameter (at 2^(nd) node):5.0 mm   Surface: Pubescent Leaf:   Shape: Ovate   Lobes: Lobed  Length: 15.0 cm   Width: 13.0 cm   Size: Longer than wide   Pubescence(both upper and lower surfaces): Pubescent   Color:     Upper surface:RHS 137B     Lower surface: RHS 137C Flower:   Staminate flowers perplant at first fruit set: 8   Diameter across staminate: 3.5 cm   Color:RHS 4A with slight RHS 144A Fruit:   General fruit type: Oblong   Maturefruit shape: Oval   Length: 14.0 cm   Diameter at midsection: 12.0 cm  Average weight: 823 g   Maximum weight: 1167 g   Surface: Smooth  Skin color pattern: Stripe   Primary color: RHS 146D   Secondarycolor: RHS 139A Rind:   Texture: Brittle   Penetrometer reading: 0.96 lb  Thickness (blossom end): 1.0 mm   Thickness (sides): 2.0 mm Flesh:  Texture: Soft   Courseness: Course fibrous   Color: RHS 6A (yellow)Seed:   Size: Medium   Length: 9.0 mm   Width: 6.0 mm   Thickness: 2.0mm   Number of seeds per fruit: 80   Color: RHS 200A

Disease resistance: Not tested

This invention is also directed to methods for producing a watermelonplant by crossing a first parent watermelon plant with a second parentwatermelon plant, wherein the first parent watermelon plant or secondparent watermelon plant is the watermelon plant from watermelon lineCA9. Further, both the first parent watermelon plant and second parentwatermelon plant may be from watermelon line CA9. Therefore, any methodsusing watermelon line CA9 are part of this invention: selfing,backcrosses, hybrid breeding, and crosses to populations. Any plantsproduced using watermelon line CA9 as at least one parent are within thescope of this invention.

Additional methods include, but are not limited to, expression vectorsintroduced into plant tissues using a direct gene transfer method, suchas microprojectile-mediated delivery, DNA injection, electroporation,and the like. More preferably, expression vectors are introduced intoplant tissues by using either microprojectile-mediated delivery with abiolistic device or by using Agrobacterium-mediated transformation.Transformed plants obtained with the protoplasm of the invention areintended to be within the scope of this invention.

Further Embodiments of the Invention

Watermelon in general is an important and valuable vegetable crop. Thus,a continuing goal of watermelon plant breeders is to develop stable,high yielding watermelon cultivars that are agronomically sound. Toaccomplish this goal, the watermelon breeder must select and developwatermelon plants with traits that result in superior cultivars.

Proper testing should detect any major faults and establish the level ofsuperiority or improvement over current cultivars. In addition toshowing superior performance, there must be a demand for a new cultivarthat is compatible with industry standards or which creates a newmarket. The introduction of a new cultivar will incur additional coststo the seed producer, the grower, processor and consumer for specialadvertising and marketing, altered seed and commercial productionpractices, and new product utilization. The testing preceding release ofa new cultivar should take into consideration research and developmentcosts, as well as technical superiority of the final cultivar. Forseed-propagated cultivars, it must be feasible to produce seed easilyand economically.

Choice of breeding or selection methods depends on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of cultivar used commercially (e.g., F₁ hybrid cultivar, purelinecultivar, etc.). For highly heritable traits, a choice of superiorindividual plants evaluated at a single location will be effective,whereas for traits with low heritability, selection should be based onmean values obtained from replicated evaluations of families of relatedplants. Popular selection methods commonly include pedigree selection,modified pedigree selection, mass selection, and recurrent selection.

The complexity of inheritance influences choice of the breeding method.Backcross breeding is used to transfer one or a few favorable genes fora highly heritable trait into a desirable cultivar. This approach hasbeen used extensively for breeding disease-resistant cultivars. Variousrecurrent selection techniques are used to improve quantitativelyinherited traits controlled by numerous genes. The use of recurrentselection in self-pollinating crops depends on the ease of pollination,the frequency of successful hybrids from each pollination, and thenumber of hybrid offspring from each successful cross.

Each breeding program should include a periodic, objective evaluation ofthe efficiency of the breeding procedure. Evaluation criteria varydepending on the goal and objectives, but should include gain fromselection per year based on comparisons to an appropriate standard, theoverall value of the advanced breeding lines, and the number ofsuccessful cultivars produced per unit of input (e.g., per year, perdollar expended, etc.).

Promising advanced breeding lines are thoroughly tested and compared toappropriate standards in environments representative of the commercialtarget area(s) for at least three years. The best lines are candidatesfor new commercial cultivars. Those still deficient in a few traits areused as parents to produce new populations for further selection.

These processes, which lead to the final step of marketing anddistribution, usually take from ten to twenty years from the time thefirst cross or selection is made. Therefore, development of newcultivars is a time-consuming process that requires precise forwardplanning, efficient use of resources, and a minimum of changes indirection.

A most difficult task is the identification of individuals that aregenetically superior, because for most traits the true genotypic valueis masked by other confounding plant traits or environmental factors.One method of identifying a superior plant is to observe its performancerelative to other experimental plants and to a widely grown standardcultivar. If a single observation is inconclusive, replicatedobservations provide a better estimate of its genetic worth.

The goal of watermelon plant breeding is to develop new, unique, andsuperior watermelon cultivars and hybrids. The breeder initially selectsand crosses two or more parental lines, followed by repeated selfing andselection, producing many new genetic combinations. The breeder cantheoretically generate billions of different genetic combinations viacrossing, selfing, and mutations. The breeder has no direct control atthe cellular level. Therefore, two breeders will never develop the sameline, or even very similar lines, having the same watermelon traits.

Each year, the plant breeder selects the germplasm to advance to thenext generation. This germplasm is grown under different geographical,climatic, and soil conditions, and further selections are then madeduring, and at the end of, the growing season. The cultivars that aredeveloped are unpredictable. This unpredictability is because thebreeder's selection occurs in unique environments, with no control atthe DNA level (using conventional breeding procedures), and withmillions of different possible genetic combinations being generated. Abreeder of ordinary skill in the art cannot predict the final resultinglines he develops, except possibly in a very gross and general fashion.The same breeder cannot produce the same line twice by using the exactsame original parents and the same selection techniques. Thisunpredictability results in the expenditure of large research monies todevelop superior watermelon cultivars.

The development of commercial watermelon cultivars requires thedevelopment of watermelon varieties, the crossing of these varieties andselection of superior hybrid crosses. The hybrid seed is produced bymanual crosses between selected male-fertile parents or by using malesterility systems. These hybrids are selected for certain single genetraits such as fruit color, flower color, pubescence color or herbicideresistance which indicate that the seed is truly a hybrid. Additionaldata on parental lines, as well as the phenotype of the hybrid,influence the breeder's decision whether to continue with the specifichybrid cross.

Pedigree breeding is used commonly for the improvement ofself-pollinating crops or inbred lines of cross-pollinating crops. Twoparents which possess favorable, complementary traits are crossed toproduce an F₁. An F₂ population is produced by selfing one or severalF₁'s or by intercrossing two F₁'s (sib mating). Selection of the bestindividuals is usually begun in the F₂ population. Then, beginning inthe F₃, the best individuals in the best families are selected.Replicated testing of families, or hybrid combinations involvingindividuals of these families, often follows in the F₄ generation toimprove the effectiveness of selection for traits with low heritability.At an advanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines are tested for potentialrelease as new cultivars.

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified or createdby intercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued.

Backcross breeding has been used to transfer genes for a simplyinherited, highly heritable trait into a desirable homozygous cultivaror line that is the recurrent parent. The source of the trait to betransferred is called the donor parent. The resulting plant is expectedto have the attributes of the recurrent parent (e.g., cultivar) and thedesirable trait transferred from the donor parent. After the initialcross, individuals possessing the phenotype of the donor parent areselected and repeatedly crossed (backcrossed) to the recurrent parent.The resulting plant is expected to have the attributes of the recurrentparent (e.g., cultivar) and the desirable trait transferred from thedonor parent.

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F₂ to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F₂ individuals. The number of plants in a population declineswith each generation due to failure of some seeds to germinate or someplants to produce at least one seed. As a result, not all of the F₂plants originally sampled in the population will be represented by aprogeny when generation advance is completed.

In a multiple-seed procedure, watermelon breeders commonly harvest twoor more seeds from the fruit of each plant in a population and bulk themto form a bulk sample. Part of the bulk is used to plant the nextgeneration and part is put in reserve. The procedure has been referredto as modified single-seed descent or the “pod-bulk” (for bean crops)technique.

The multiple-seed procedure has been used to save labor at harvest. Itis considerably faster to extract seeds with a machine than to removeone seed from each by hand for the single-seed procedure. Themultiple-seed procedure also makes it possible to plant the same numberof seeds of a population each generation of inbreeding. Enough seeds areharvested to make up for those plants that did not germinate or produceseed.

In addition to phenotypic observations, the genotype of a plant can alsobe examined. There are many laboratory-based techniques available forthe analysis, comparison and characterization of plant genotype; amongthese are Isozyme Electrophoresis, Restriction Fragment LengthPolymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs),Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Amplified Fragment Length polymorphisms (AFLPs), Simple Sequence Repeats(SSRs—which are also referred to as Microsatellites), and SingleNucleotide Polymorphisms (SNPs).

Isozyme Electrophoresis and RFLPs have been widely used to determinegenetic composition. Shoemaker and Olsen (Molecular Linkage Map ofSoybean (Glycine max), pp. 6.131-6.138 in S.J. O′Brien (ed.) GeneticMaps: Locus Maps of Complex Genomes, Cold Spring Harbor LaboratoryPress, Cold Spring Harbor, NY (1993)) developed a molecular geneticlinkage map that consisted of 25 linkage groups with about 365 RFLP, 11RAPD, three classical markers, and four isozyme loci. See also,Shoemaker, R.C., RFLP Map of Soybean, pp. 299-309, in Phillips, R.L. andVasil, I.K. (eds.), DNA-Based Markers in Plants, Kluwer Academic Press,Dordrecht, the Netherlands (1994).

SSR technology is currently the most efficient and practical markertechnology; more marker loci can be routinely used and more alleles permarker locus can be found using SSRs in comparison to RFLPs. Forexample, Diwan and Cregan described a highly polymorphic microsatellitelocus in soybean with as many as 26 alleles. Diwan, N. and Cregan, P.B., Theon. Appl. Genet., 95:22-225 (1997). SNPs may also be used toidentify the unique genetic composition of the invention and progenyvarieties retaining that unique genetic composition. Various molecularmarker techniques may be used in combination to enhance overallresolution.

Molecular markers, which include markers identified through the use oftechniques such as Isozyme Electrophoresis, RFLPs, RAPDs, AP-PCR, DAF,SCARs, AFLPs, SSRs, and SNPs, may be used in plant breeding. One use ofmolecular markers is Quantitative Trait Loci (QTL) mapping. QTL mappingis the use of markers which are known to be closely linked to allelesthat have measurable effects on a quantitative trait. Selection in thebreeding process is based upon the accumulation of markers linked to thepositive effecting alleles and/or the elimination of the markers linkedto the negative effecting alleles from the plant's genome.

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select toward the genome of the recurrent parent and against themarkers of the donor parent. This procedure attempts to minimize theamount of genome from the donor parent that remains in the selectedplants. It can also be used to reduce the number of crosses back to therecurrent parent needed in a backcrossing program. The use of molecularmarkers in the selection process is often called genetic marker enhancedselection or marker-assisted selection. Molecular markers may also beused to identify and exclude certain sources of germplasm as parentalvarieties or ancestors of a plant by providing a means of trackinggenetic profiles through crosses.

Mutation breeding is another method of introducing new traits intowatermelon varieties. Mutations that occur spontaneously or areartificially induced can be useful sources of variability for a plantbreeder. The goal of artificial mutagenesis is to increase the rate ofmutation for a desired characteristic. Mutation rates can be increasedby many different means including temperature, long-term seed storage,tissue culture conditions, radiation (such as X-rays, Gamma rays,neutrons, Beta radiation, or ultraviolet radiation), chemical mutagens(such as base analogs like 5-bromo-uracil), antibiotics, alkylatingagents (such as sulfur mustards, nitrogen mustards, epoxides,ethyleneamines, sulfates, sulfonates, sulfones, or lactones), azide,hydroxylamine, nitrous acid, or acridines. Once a desired trait isobserved through mutagenesis the trait may then be incorporated intoexisting germplasm by traditional breeding techniques. Details ofmutation breeding can be found in Principles of Cultivar Development byFehr, Macmillan Publishing Company (1993).

The production of double haploids can also be used for the developmentof homozygous varieties in a breeding program. Double haploids areproduced by the doubling of a set of chromosomes from a heterozygousplant to produce a completely homozygous individual. For example, seeWan, et al., Theor. Appl. Genet., 77:889-892 (1989).

Descriptions of other breeding methods that are commonly used fordifferent traits and crops can be found in one of several referencebooks (e.g., Principles of Plant Breeding, John Wiley and Son, pp.115-161 (1960); Allard (1960); Simmonds (1979); Sneep, et al. (1979);Fehr (1987); “Carrots and Related Vegetable Umbelliferae,” Rubatzky,V.E., et al. (1999).

With the advent of molecular biological techniques that have allowed theisolation and characterization of genes that encode specific proteinproducts, scientists in the field of plant biology developed a stronginterest in engineering the genome of plants to contain and expressforeign genes, or additional, or modified versions of native, orendogenous, genes (perhaps driven by different promoters) in order toalter the traits of a plant in a specific manner. Any DNA sequences,whether from a different species or from the same species, which areintroduced into the genome using transformation or various breedingmethods, are referred to herein collectively as “transgenes.” Over thelast fifteen to twenty years, several methods for producing transgenicplants have been developed, and the present invention, in particularembodiments, also relates to transformed versions of the claimed line.

Nucleic acids or polynucleotides refer to RNA or DNA that is linear orbranched, single or double stranded, or a hybrid thereof. The term alsoencompasses RNA/DNA hybrids. These terms also encompass untranslatedsequence located at both the 3′ and 5′ ends of the coding region of thegene: at least about 1000 nucleotides of sequence upstream from the 5′end of the coding region and at least about 200 nucleotides of sequencedownstream from the 3′ end of the coding region of the gene. Less commonbases, such as inosine, 5-methylcytosine, 6-methyladenine, hypoxanthine,and others can also be used for antisense, dsRNA, and ribozyme pairing.For example, polynucleotides that contain C-5 propyne analogues ofuridine and cytidine have been shown to bind RNA with high affinity andto be potent antisense inhibitors of gene expression. Othermodifications, such as modification to the phosphodiester backbone, orthe 2′-hydroxy in the ribose sugar group of the RNA can also be made.The antisense polynucleotides and ribozymes can consist entirely ofribonucleotides, or can contain mixed ribonucleotides anddeoxyribonucleotides. The polynucleotides of the invention may beproduced by any means, including genomic preparations, cDNApreparations, in vitro synthesis, RT-PCR, and in vitro or in vivotranscription.

Plant transformation involves the construction of an expression vectorthat will function in plant cells. Such a vector comprises DNAcomprising a gene under control of, or operatively linked to, aregulatory element (for example, a promoter). The expression vector maycontain one or more such operably linked gene/regulatory elementcombinations. The vector(s) may be in the form of a plasmid, and can beused alone or in combination with other plasmids, to provide transformedwatermelon plants using transformation methods as described below toincorporate transgenes into the genetic material of the watermelonplant(s). Expression Vectors for Watermelon Transformation: Marker Genes

Expression vectors include at least one genetic marker, operably linkedto a regulatory element (for example, a promoter) that allowstransformed cells containing the marker to be either recovered bynegative selection, i e , inhibiting growth of cells that do not containthe selectable marker gene, or by positive selection, i.e., screeningfor the product encoded by the genetic marker. Many commonly usedselectable marker genes for plant transformation are well known in thetransformation arts, and include, for example, genes that code forenzymes that metabolically detoxify a selective chemical agent which maybe an antibiotic or an herbicide, or genes that encode an altered targetwhich is insensitive to the inhibitor. A few positive selection methodsare also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptII) gene, isolated from transposonTn5, which when placed under the control of plant regulatory signalsconfers resistance to kanamycin. Fraley, et al., PNAS, 80:4803 (1983).Another commonly used selectable marker gene is the hygromycinphosphotransferase gene which confers resistance to the antibiotichygromycin. Vanden Elzen, et al., Plant Mol. Biol., 5:299 (1985).

Additional selectable marker genes of bacterial origin that conferresistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase, aminoglycoside-3′-adenyl transferase,the bleomycin resistance determinant. Hayford, et al., Plant Physiol.,86:1216 (1988); Jones, et al., Mol. Gen. Genet., 210:86 (1987); Svab, etal., Plant Mol. Biol., 14:197 (1990); Hille, et al., Plant Mol. Biol.,7:171 (1986). Other selectable marker genes confer resistance toherbicides such as glyphosate, glufosinate, or bromoxynil. Comai, etal., Nature, 317:741-744 (1985); Gordon-Kamm, et al., Plant Cell,2:603-618 (1990); and Stalker, et al., Science, 242:419-423 (1988).

Selectable marker genes for plant transformation that are not ofbacterial origin include, for example, mouse dihydrofolate reductase,plant 5-enolpyruvylshikimate-3-phosphate synthase, and plantacetolactate synthase. Eichholtz, et al., Somatic Cell Mol. Genet.,13:67 (1987); Shah, et al., Science, 233:478 (1986); and Charest, etal., Plant Cell Rep., 8:643 (1990).

Another class of marker genes for plant transformation requiresscreening of presumptively transformed plant cells rather than directgenetic selection of transformed cells for resistance to a toxicsubstance such as an antibiotic. These genes are particularly useful toquantify or visualize the spatial pattern of expression of a gene inspecific tissues and are frequently referred to as reporter genesbecause they can be fused to a gene or gene regulatory sequence for theinvestigation of gene expression. Commonly used genes for screeningpresumptively transformed cells include α-glucuronidase (GUS),α-galactosidase, luciferase and chloramphenicol, acetyltransferase.Jefferson, R. A., Plant Mol. Biol., 5:387 (1987); Teeri, et al., EMBOJ., 8:343 (1989); Koncz, et al., PNAS, 84:131 (1987); and DeBlock, etal., EMBO J., 3:1681 (1984).

In vivo methods for visualizing GUS activity that do not requiredestruction of plant tissues are available. Molecular Probes,Publication 2908, IMAGENE GREEN, pp. 1-4 (1993) and Naleway, et al., J.Cell Biol., 115:151a (1991). However, these in vivo methods forvisualizing GUS activity have not proven useful for recovery oftransformed cells because of low sensitivity, high fluorescentbackgrounds, and limitations associated with the use of luciferase genesas selectable markers.

More recently, a gene encoding Green Fluorescent Protein (GFP) has beenutilized as a marker for gene expression in prokaryotic and eukaryoticcells. Chalfie, et al., Science, 263:802 (1994). GFP and mutants of GFPmay be used as screenable markers.

Expression Vectors for Watermelon Transformation: Promoters

Genes included in expression vectors must be driven by a nucleotidesequence comprising a regulatory element (for example, a promoter).Several types of promoters are now well known in the transformationarts, as are other regulatory elements that can be used alone or incombination with promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred.”Promoters which initiate transcription only in certain tissue arereferred to as “tissue-specific.” A “cell type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter which is under environmental control. Examples ofenvironmental conditions that may effect transcription by induciblepromoters include anaerobic conditions or the presence of light.Tissue-specific, tissue-preferred, cell type specific, and induciblepromoters constitute the class of “non-constitutive” promoters. A“constitutive” promoter is a promoter which is active under mostenvironmental conditions.

A. Inducible Promoters:

An inducible promoter is operably linked to a gene for expression inwatermelon. Optionally, the inducible promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in watermelon. With an inducible promoter, therate of transcription increases in response to an inducing agent.

Any inducible promoter can be used in the instant invention. See Ward,et al., Plant Mol. Biol., 22:361-366 (1993). Exemplary induciblepromoters include, but are not limited to, that from the ACEI systemwhich responds to copper (Meft, et al., PNAS, 90:4567-4571 (1993)); In2gene from maize which responds to benzenesulfonamide herbicide safeners(Hershey, et al., Mol. Gen. Genet., 227:229-237 (1991) and Gatz, et al.,Mol. Gen. Genet., 243:32-38 (1994)) or Tet repressor from TnlO (Gatz, etal., Mol. Gen. Genet., 227:229-237 (1991)). A particularly preferredinducible promoter is a promoter that responds to an inducing agent towhich plants do not normally respond. An exemplary inducible promoter isthe inducible promoter from a steroid hormone gene, the transcriptionalactivity of which is induced by a glucocorticosteroid hormone. Schena,et al., PNAS, 88:0421 (1991).

B. Constitutive Promoters:

A constitutive promoter is operably linked to a gene for expression inwatermelon or the constitutive promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in watermelon.

Many different constitutive promoters can be utilized in the instantinvention. Exemplary constitutive promoters include, but are not limitedto, the promoters from plant viruses such as the 35S promoter from CaMV(Odell, et al., Nature, 313:810-812 (1985)) and the promoters from suchgenes as rice actin (McElroy, et al., Plant Cell, 2:163-171 (1990));ubiquitin (Christensen, et al., Plant Mol. Biol., 12:619-632 (1989) andChristensen, et al., Plant Mol. Biol., 18:675-689 (1992)); pEMU (Last,et al., Theor. Appl. Genet., 81:581-588 (1991)); MAS (Velten, et al.,EMBO J., 3:2723-2730 (1984)) and maize H3 histone (Lepetit, et al., Mol.Gen. Genet., 231:276-285 (1992) and Atanassova, et al., Plant J., 2(3):291-300 (1992)). The ALS promoter, Xbal/Ncol fragment 5′ to theBrassica napus ALS3 structural gene (or a nucleotide sequence similarityto said Xbal/Ncol fragment), represents a particularly usefulconstitutive promoter. See PCT Application No. WO 96/30530.

C. Tissue-Specific or Tissue-Preferred Promoters:

A tissue-specific promoter is operably linked to a gene for expressionin watermelon. Optionally, the tissue-specific promoter is operablylinked to a nucleotide sequence encoding a signal sequence which isoperably linked to a gene for expression in watermelon. Plantstransformed with a gene of interest operably linked to a tissue-specificpromoter produce the protein product of the transgene exclusively, orpreferentially, in a specific tissue.

Any tissue-specific or tissue-preferred promoter can be utilized in theinstant invention. Exemplary tissue-specific or tissue-preferredpromoters include, but are not limited to, a root-preferred promoter,such as that from the phaseolin gene (Murai, et al., Science, 23:476-482(1983) and Sengupta-Gopalan, et al., PNAS, 82:3320-3324 (1985)); aleaf-specific and light-induced promoter such as that from cab orrubisco (Simpson, et al., EMBO J., 4(11):2723-2729 (1985) and Timko, etal., Nature, 318:579-582 (1985)); an anther-specific promoter such asthat from LAT52 (Twell, et al., Mol. Gen. Genet., 217:240-245 (1989)); apollen-specific promoter such as that from Zml3 (Guerrero, et al., Mol.Gen. Genet., 244:161-168 (1993)) or a microspore-preferred promoter suchas that from apg (Twell, et al., Sex. Plant Reprod., 6:217-224 (1993)).

Signal Sequences for Targeting Proteins to Subcellular Compartments

Transport of protein produced by transgenes to a subcellular compartmentsuch as the chloroplast, vacuole, peroxisome, glyoxysome, cell wall, ormitochondrion, or for secretion into the apoplast, is accomplished bymeans of operably linking the nucleotide sequence encoding a signalsequence to the 5′ and/or 3′ region of a gene encoding the protein ofinterest. Targeting sequences at the 5′ and/or 3′ end of the structuralgene may determine, during protein synthesis and processing, where theencoded protein is ultimately compartmentalized.

The presence of a signal sequence directs a polypeptide to either anintracellular organelle or subcellular compartment or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample, Becker, et al., Plant Mol. Biol., 20:49 (1992); Close, P. S.,Master's Thesis, Iowa State University (1993); Knox, C., et al.,“Structure and Organization of Two Divergent Alpha-Amylase Genes fromBarley,” Plant Mol. Biol., 9:3-17 (1987); Lerner, et al., PlantPhysiol., 91:124-129 (1989); Fontes, et al., Plant Cell, 3:483-496(1991); Matsuoka, et al., PNAS, 88:834 (1991); Gould, et al., J. Cell.Biol., 108:1657 (1989); Creissen, et al., Plant J., 2:129 (1991);Kalderon, et al., A short amino acid sequence able to specify nuclearlocation, Cell, 39:499-509 (1984); and Steifel, et al., Expression of amaize cell wall hydroxyproline-rich glycoprotein gene in early leaf androot vascular differentiation, Plant Cell, 2:785-793 (1990).

Foreign Protein Genes and Agronomic Genes

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, Anal. Biochem., 114:92-6(1981).

According to a preferred embodiment, the transgenic plant provided forcommercial production of foreign protein is watermelon. In anotherpreferred embodiment, the biomass of interest is seed. For therelatively small number of transgenic plants that show higher levels ofexpression, a genetic map can be generated, primarily via conventionalRFLP, PCR, and SSR analysis, which identifies the approximatechromosomal location of the integrated DNA molecule. For exemplarymethodologies in this regard, see Methods in Plant Molecular Biology andBiotechnology, Glick and Thompson Eds., 269:284, CRC Press, Boca Raton(1993). Map information concerning chromosomal location is useful forproprietary protection of a subject transgenic plant. If unauthorizedpropagation is undertaken and crosses made with other germplasm, the mapof the integration region can be compared to similar maps for suspectplants, to determine if the latter have a common parentage with thesubject plant. Map comparisons would involve hybridizations, RFLP, PCR,SSR, and sequencing, all of which are conventional techniques.

Likewise, by means of the present invention, agronomic genes can beexpressed in transformed plants. More particularly, plants can begenetically engineered to express various phenotypes of agronomicinterest. Exemplary genes implicated in this regard include, but are notlimited to, those categorized below:

A. Genes That Confer Resistance to Pests or Disease and That Encode:

1. Plant disease resistance genes. Plant defenses are often activated byspecific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant line can be transformed with a clonedresistance gene to engineer plants that are resistant to specificpathogen strains. See, for example, Jones, et al., Science, 266:789(1994) (cloning of the tomato Cf-9 gene for resistance to Cladosporiumfulvum); Martin, et al., Science, 262:1432 (1993) (tomato Pto gene forresistance to Pseudomonas syringae pv. tomato encodes a protein kinase);and Mindrinos, et al., Cell, 78:1089 (1994) (Arabidopsis RSP2 gene forresistance to Pseudomonas syringae).

2. A Bacillus thuringiensis protein, a derivative thereof, or asynthetic polypeptide modeled thereon. See, for example, Geiser, et al.,Gene, 48:109 (1986), who disclose the cloning and nucleotide sequence ofa Bt δ-endotoxin gene. Moreover, DNA molecules encoding δ-endotoxingenes can be purchased from American Type Culture Collection, Manassas,Virginia, for example, under ATCC Accession Nos. 40098, 67136, 31995,and 31998.

3. A lectin. See, for example, the disclosure by Van Damme, et al.,Plant Mol. Biol., 24:25 (1994), who disclose the nucleotide sequences ofseveral Clivia miniata mannose-binding lectin genes.

4. A vitamin-binding protein such as avidin. See PCT Application No. US93/06487, the contents of which are hereby incorporated by reference.The application teaches the use of avidin and avidin homologues aslarvicides against insect pests.

5. An enzyme inhibitor, for example, a protease or proteinase inhibitor,or an amylase inhibitor. See, for example, Abe, et al., J. Biol. Chem.,262:16793 (1987) (nucleotide sequence of rice cysteine proteinaseinhibitor); Huub, et al., Plant Mol. Biol., 21:985 (1993) (nucleotidesequence of cDNA encoding tobacco proteinase inhibitor I); and Sumitani,et al., Biosci. Biotech. Biochem., 57:1243 (1993) (nucleotide sequenceof Streptomyces nitrosporeus α-amylase inhibitor).

6. An insect-specific hormone or pheromone, such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock, et al., Nature, 344:458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

7. An insect-specific peptide or neuropeptide which, upon expression,disrupts the physiology of the affected pest. For example, see thedisclosures of Regan, J. Biol. Chem., 269:9 (1994) (expression cloningyields DNA coding for insect diuretic hormone receptor) and Pratt, etal., Biochem. Biophys. Res. Comm., 163:1243 (1989) (an allostatin isidentified in Diploptera puntata). See also, U.S. Pat. No. 5,266,317 toTomalski, et al., who disclose genes encoding insect-specific, paralyticneurotoxins.

8. An insect-specific venom produced in nature, by a snake, a wasp, etc.For example, see Pang, et al., Gene, 116:165 (1992), for disclosure ofheterologous expression in plants of a gene coding for a scorpioninsectotoxic peptide.

9. An enzyme responsible for a hyper-accumulation of a monoterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivative,or another non-protein molecule with insecticidal activity.

10. An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase, and a glucanase, whether natural or synthetic. See PCTApplication No. WO 93/02197 in the name of Scott, et al., whichdiscloses the nucleotide sequence of a callase gene. DNA molecules whichcontain chitinase-encoding sequences can be obtained, for example, fromthe ATCC under Accession Nos. 39637 and 67152. See also, Kramer, et al.,Insect Biochem. Mol. Biol., 23:691 (1993), who teach the nucleotidesequence of a cDNA encoding tobacco hornworm chitinase, and Kawalleck,et al., Plant Mol. Biol., 21:673 (1993), who provide the nucleotidesequence of the parsley ubi4-2 polyubiquitin gene.

11. A molecule that stimulates signal transduction. For example, see thedisclosure by Botella, et al., Plant Mol. Biol., 24:757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess,et al., Plant Physiol., 104:1467 (1994), who provide the nucleotidesequence of a maize calmodulin cDNA clone.

12. A hydrophobic moment peptide. See PCT Application No. WO 95/16776(disclosure of peptide derivatives of tachyplesin which inhibit fungalplant pathogens) and PCT

Application No. WO 95/18855 (teaches synthetic antimicrobial peptidesthat confer disease resistance), the respective contents of which arehereby incorporated by reference.

13. A membrane permease, a channel former, or a channel blocker. Forexample, see the disclosure of Jaynes, et al., Plant Sci., 89:43 (1993),of heterologous expression of a cecropin-β, lytic peptide analog torender transgenic tobacco plants resistant to Pseudomonas solanacearum.

14. A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy, et al., Ann. Rev. Phytopathol.,28:451 (1990). Coat protein-mediated resistance has been conferred upontransformed plants against alfalfa mosaic virus, cucumber mosaic virus,tobacco streak virus, potato virus X, potato virus Y, tobacco etchvirus, tobacco rattle virus, and tobacco mosaic virus. Id.

15. An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. SeeTaylor, et al., Abstract #497, Seventh Int'l Symposium on MolecularPlant-Microbe Interactions, Edinburgh, Scotland (1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

16. A virus-specific antibody. See, for example, Tavladoraki, et al.,Nature, 366:469 (1993), who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

17. A developmental-arrestive protein produced in nature by a pathogenor a parasite. Thus, fungal endo-α-1,4-D-polygalacturonases facilitatefungal colonization and plant nutrient released by solubilizing plantcell wall homo-α-1,4-D-galacturonase. See Lamb, et al., Bio/technology,10:1436 (1992). The cloning and characterization of a gene which encodesa bean endopolygalacturonase-inhibiting protein is described by Toubart,et al., Plant J., 2:367 (1992).

18. A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann, et al., Bio/technology, 10:305 (1992), have shownthat transgenic plants expressing the barley ribosome-inactivating genehave an increased resistance to fungal disease.

19. Genes involved in the Systemic Acquired Resistance (SAR) Responseand/or the pathogenesis-related genes. Briggs, S., Current Biology, 5(2)(1995).

20. Antifungal genes. See Cornelissen and Melchers, Plant Physiol.,101:709-712 (1993); Parijs et al., Planta 183:258-264 (1991) andBushnell et al., Can. J. of Plant Path. 20(2):137-149 (1998).

21. Genes that confer resistance to Phytophthora root rot, such as theRps 1, Rps 1-a, Rps 1-b, Rps 1-c, Rps 1-d, Rps 1-e, Rps 1-k, Rps 2, Rps3-a, Rps 3-b, Rps 3-c, Rps 4, Rps 5, Rps 6, Rps 7 and other Rps genes.See, for example, Shoemaker et al., Phytophthora Root Rot ResistanceGene Mapping in Soybean, Plant Genome IV Conference, San Diego, Calif.(1995).

Any of the above listed disease or pest resistance genes (1-21) can beintroduced into the claimed watermelon cultivar through a variety ofmeans including but not limited to transformation and crossing.

B. Genes That Confer Resistance to an Herbicide:

1. An herbicide that inhibits the growing point or meristem, such as animidazolinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee, etal., EMBO J., 7:1241 (1988) and Miki, et al., Theor. Appl. Genet.,80:449 (1990), respectively.

2. Glyphosate (resistance conferred by mutant5-enolpyruvlshikimate-3-phosphate synthase (EPSPS) and aroA genes,respectively) and other phosphono compounds, such as glufosinate(phosphinothricin acetyl transferase (PAT), dicamba and Streptomyceshygroscopicus phosphinothricin-acetyl transferase PAT bar genes), andpyridinoxy or phenoxy proprionic acids and cyclohexones (ACCaseinhibitor-encoding genes). See, for example, U.S. Pat. No. 4,940,835 toShah, et al., which discloses the nucleotide sequence of a form of EPSPSwhich can confer glyphosate resistance. A DNA molecule encoding a mutantaroA gene can be obtained under ATCC Accession No. 39256, and thenucleotide sequence of the mutant gene is disclosed in U.S. Pat. No.4,769,061 to Comai. See also, Umaballava-Mobapathie in TransgenicResearch, 8:1, 33-44 (1999) that discloses Lactuca sativa resistant toglufosinate. European Patent Application No. 0 333 033 to Kumada, etal., and U.S. Pat. No. 4,975,374 to Goodman, et al., disclose nucleotidesequences of glutamine synthetase genes which confer resistance toherbicides, such as L-phosphinothricin. The nucleotide sequence of aphosphinothricin-acetyl-transferase gene is provided in EuropeanApplication No. 0 242 246 to Leemans, et al. DeGreef, et al.,Bio/technology, 7:61 (1989), describe the production of transgenicplants that express chimeric bar genes coding for phosphinothricinacetyl transferase activity. Exemplary of genes conferring resistance tophenoxy proprionic acids and cyclohexones, such as sethoxydim andhaloxyfop, are the Accl-S1, Accl-S2, and Accl-S3 genes described byMarshall, et al., Theor. Appl. Genet., 83:435 (1992).

3. An herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+ genes) and a benzonitrile (nitrilase gene). Przibilla, et al.,Plant Cell, 3:169 (1991), describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, andDNA molecules containing these genes are available under ATCC AccessionNos. 53435, 67441, and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes, et al., Biochem. J.,285:173 (1992).

4. Acetohydroxy acid synthase, which has been found to make plants thatexpress this enzyme resistant to multiple types of herbicides, has beenintroduced into a variety of plants. See Hattori, et al., Mol. Gen.Genet., 246:419 (1995). Other genes that confer tolerance to herbicidesinclude a gene encoding a chimeric protein of rat cytochrome P4507A1 andyeast NADPH-cytochrome P450 oxidoreductase (Shiota, et al., PlantPhysiol., 106:17 (1994)), genes for glutathione reductase and superoxidedismutase (Aono, et al., Plant Cell Physiol., 36:1687 (1995)), and genesfor various phosphotransferases (Datta, et al., Plant Mol. Biol., 20:619(1992)).

5. Protoporphyrinogen oxidase (protox) is necessary for the productionof chlorophyll, which is necessary for all plant survival. The protoxenzyme serves as the target for a variety of herbicidal compounds. Theseherbicides also inhibit growth of all the different species of plantspresent, causing their total destruction. The development of plantscontaining altered protox activity which are resistant to theseherbicides are described in U.S. Pat. Nos. 6,288, 306, 6,282,837,5,767,373, and International Publication WO 01/12825.

Any of the above listed herbicide genes (1-5) can be introduced into theclaimed watermelon cultivar through a variety of means including, butnot limited to, transformation and crossing.

C. Genes That Confer or Contribute to a Value-Added Trait, such as:

1. Modified fatty acid metabolism, for example, by transforming a plantwith an antisense gene of stearyl-ACP desaturase to increase stearicacid content of the plant. See Knultzon et al., Proc. Natl. Acad. Sci.USA 89:2625 (1992).

2. Decreased phytate content—1) Introduction of a phytase-encoding geneenhances breakdown of phytate, adding more free phosphate to thetransformed plant. For example, see Van Hartingsveldt et al., Gene127:87 (1993), for a disclosure of the nucleotide sequence of anAspergillus niger phytase gene. 2) A gene could be introduced thatreduced phytate content. See Raboy et al., Maydica 35:383 (1990).

3. Increased sweetness of the watermelon by introducing a gene codingfor monellin that elicits a flavor 100,000 times sweeter than sugar on amolar basis. See Penarrubia, et al., Bio/technology, 10:561-564 (1992).

4. Modified fatty acid metabolism, for example, by introducing into aplant an antisense gene of stearyl-ACP desaturase to increase stearicacid content of the plant. See Knultzon, et al., PNAS, 89:2625 (1992).

5. Modified carbohydrate composition effected, for example, byintroducing into plants a gene coding for an enzyme that alters thebranching pattern of starch. See Shiroza, et al., J. Bacteriol., 170:810(1988) (nucleotide sequence of Streptococcus mutantsfructosyltransferase gene); Steinmetz, et al., Mol. Gen. Genet., 20:220(1985) (nucleotide sequence of Bacillus subtilis levansucrase gene);Pen, et al., Bio/technology, 10:292 (1992) (production of transgenicplants that express Bacillus lichenifonnis α-amylase); Elliot, et al.,Plant Mol. Biol., 21:515 (1993) (nucleotide sequences of tomatoinvertase genes); Sogaard, et al., J. Biol. Chem., 268:22480 (1993)(site-directed mutagenesis of barley α-amylase gene); and Fisher, etal., Plant Physiol., 102:1045 (1993) (maize endosperm starch branchingenzyme II).

6. Elevated oleic acid via FAD-2 gene modification and/or decreasedlinolenic acid via FAD-3 gene modification. See U.S. Pat. Nos.6,063,947; 6,323,392; and PCT Publication WO 93/11245.

D. Genes that Control Male-Sterility:

1. Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN-Ac-PPT. See International Publication WO 01/29237.

2. Introduction of various stamen-specific promoters. See InternationalPublications WO 92/13956 and WO 92/13957.

3. Introduction of the barnase and the barstar genes. See Paul, et al.,Plant Mol. Biol., 19:611-622 (1992).

Methods for Watermelon Transformation

Numerous methods for plant transformation have been developed, includingbiological and physical, plant transformation protocols. See, forexample, Miki, et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, Glickand Thompson Eds., CRC Press, Inc., Boca Raton, pp. 67-88 (1993). Inaddition, expression vectors and in vitro culture methods for plant cellor tissue transformation and regeneration of plants are available. See,for example, Gruber, et al., “Vectors for Plant Transformation” inMethods in Plant Molecular Biology and Biotechnology, Glick and ThompsonEds., CRC Press, Inc., Boca Raton, pp. 89-119 (1993).

A. Agrobacterium-Mediated Transformation:

One method for introducing an expression vector into plants is based onthe natural transformation system of Agrobacterium. See, for example,Horsch, et al., Science, 227:1229 (1985); Curtis, et al., Journal ofExperimental Botany, 45:279, 1441-1449 (1994); Tones, et al., Plant CellTissue and Organ Culture, 34:3, 279-285 (1993); and Dinant, et al.,Molecular Breeding, 3:1, 75-86 (1997). A. tumefaciens and A. rhizogenesare plant pathogenic soil bacteria which genetically transform plantcells. The Ti and Ri plasmids of A. tumefaciens and A. rhizogenes,respectively, carry genes responsible for genetic transformation of theplant. See, for example, Kado, C. I., Crit. Rev. Plant Sci., 10:1(1991). Descriptions of Agrobacterium vector systems and methods forAgrobacterium-mediated gene transfer are provided by Gruber, et al.,supra, Miki, et al., supra, and Moloney, et al., Plant Cell Rep., 8:238(1989). See also, U.S. Pat. No. 5,591,616 issued Jan. 7, 1997.

B. Direct Gene Transfer:

Several methods of plant transformation collectively referred to asdirect gene transfer have been developed as an alternative toAgrobacterium-mediated transformation. A generally applicable method ofplant transformation is microproj ectile-mediated transformation whereinDNA is carried on the surface of microprojectiles measuring 1 μm to 4μm. The expression vector is introduced into plant tissues with abiolistic device that accelerates the microprojectiles to speeds of 300m/s to 600 m/s which is sufficient to penetrate plant cell walls andmembranes. Russell, D. R., et al., Plant Cell Rep., 12 (3, January),165-169 (1993); Aragao, F. J. L., et al., Plant Mol. Biol., 20 (2,October), 357-359 (1992); Aragao, F. J. L., et al., Plant Cell Rep., 12(9, July), 483-490 (1993); Aragao, Theor. Appl. Genet., 93:142-150(1996); Kim, J., Minamikawa, T., Plant Sci., 117:131-138 (1996);Sanford, et al., Part. Sci. Technol., 5:27 (1987); Sanford, J. C.,Trends Biotech., 6:299 (1988); Klein, et al., Bio/technology, 6:559-563(1988); Sanford, J. C., Physiol. Plant, 7:206 (1990); Klein, et al.,Bio/technology, 10:268 (1992).

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang, et al., Bio/technology, 9:996 (1991).Alternatively, liposome and spheroplast SWD8732 have been used tointroduce expression vectors into plants. Deshayes, et al., EMBO J.,4:2731 (1985) and Christou, et al., PNAS, 84:3962 (1987). Direct uptakeof DNA into protoplasts using CaCl₂ precipitation, polyvinyl alcohol, orpoly-L-ornithine has also been reported. Hain, et al., Mol. Gen. Genet.,199:161 (1985) and Draper, et al., Plant Cell Physiol., 23:451 (1982).Electroporation of protoplasts and whole cells and tissues have alsobeen described. Saker, M., Kuhne, T., Biologia Plantarum, 40(4):507-514(1997/98); Donn, et al., In Abstracts of VIIth International Congress onPlant Cell and Tissue Culture IAPTC, A2-38, p. 53 (1990); D′Halluin, etal., Plant Cell, 4:1495-1505 (1992); and Spencer, et al., Plant Mol.Biol., 24:51-61 (1994). See also Chupean, et al., Bio/technology, 7:5,503-508 (1989).

Additional methods include, but are not limited to, expression vectorsintroduced into plant tissues using a direct gene transfer method, suchas microprojectile-mediated delivery, DNA injection, electroporation,and the like. More preferably, expression vectors are introduced intoplant tissues by using either microprojectile-mediated delivery with abiolistic device or by using Agrobacterium-mediated transformation.Transformed plants obtained with the protoplasm of the invention areintended to be within the scope of this invention.

Following transformation of watermelon target tissues, expression of theabove-described selectable marker genes allows for preferentialselection of transformed cells, tissues and/or plants, usingregeneration and selection methods now well known in the art.

The foregoing methods for transformation would typically be used forproducing a transgenic line. The transgenic line could then be crossedwith another (non-transformed or transformed) line in order to produce anew transgenic watermelon pollenizer. Alternatively, a genetic traitwhich has been engineered into a particular watermelon cultivar usingthe foregoing transformation techniques could be introduced into anotherline using traditional backcrossing techniques that are well known inthe plant breeding arts. For example, a backcrossing approach could beused to move an engineered trait from a public, non-elite inbred lineinto an elite inbred line, or from an inbred line containing a foreigngene in its genome into an inbred line or lines which do not containthat gene. As used herein, “crossing” can refer to a simple X by Ycross, or the process of backcrossing, depending on the context.

Gene Conversions

When the term “watermelon plant” is used in the context of the presentinvention, this also includes any gene conversions of that variety. Theterm “gene converted plant” as used herein refers to those watermelonplants which are developed by backcrossing, genetic engineering, ormutation, wherein essentially all of the desired morphological andphysiological characteristics of a variety are recovered in addition tothe one or more genes transferred into the variety via the backcrossingtechnique, genetic engineering, or mutation. Backcrossing methods can beused with the present invention to improve or introduce a characteristicinto the variety. The term “backcrossing” as used herein refers to therepeated crossing of a hybrid progeny back to the recurrent parent,i.e., backcrossing 1, 2, 3, 4, 5, 6, 7, 8, 9, or more times to therecurrent parent. The parental watermelon plant which contributes thegene for the desired characteristic is termed the “nonrecurrent” or“donor parent.” This terminology refers to the fact that thenonrecurrent parent is used one time in the backcross protocol andtherefore does not recur. The parental watermelon plant to which thegene or genes from the nonrecurrent parent are transferred is known asthe recurrent parent as it is used for several rounds in thebackcrossing protocol. Poehlman & Sleper (1994) and Fehr (1993). In atypical backcross protocol, the original variety of interest (recurrentparent) is crossed to a second variety (nonrecurrent parent) thatcarries the gene of interest to be transferred. The resulting progenyfrom this cross are then crossed again to the recurrent parent and theprocess is repeated until a watermelon plant is obtained whereinessentially all of the desired morphological and physiologicalcharacteristics of the recurrent parent are recovered in the convertedplant, in addition to the transferred gene from the nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a trait or characteristic in the original line.To accomplish this, a gene of the recurrent cultivar is modified orsubstituted with the desired gene from the nonrecurrent parent, whileretaining essentially all of the rest of the desired genetic, andtherefore the desired physiological and morphological characteristics,watermelon line CA9 of the original line. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross. One ofthe major purposes is to add some commercially desirable, agronomicallyimportant trait to the plant. The exact backcrossing protocol willdepend on the characteristic or trait being altered to determine anappropriate testing protocol. Although backcrossing methods aresimplified when the characteristic being transferred is a dominantallele, a recessive allele may also be transferred. In this instance itmay be necessary to introduce a test of the progeny to determine if thedesired characteristic has been successfully transferred.

Many gene traits have been identified that are not regularly selected inthe development of a new line but that can be improved by backcrossingtechniques. Gene traits may or may not be transgenic. Examples of thesetraits include, but are not limited to, male sterility, modified fattyacid metabolism, modified carbohydrate metabolism, herbicide resistance,resistance for bacterial, fungal, or viral disease, insect resistance,enhanced nutritional quality, industrial usage, yield stability, andyield enhancement. These genes are generally inherited through thenucleus. Several of these gene traits are described in U.S. Pat. Nos.5,777,196, 5,948,957, and 5,969,212, the disclosures of which arespecifically hereby incorporated by reference.

Tissue Culture

Further reproduction of the variety can occur by tissue culture andregeneration. Tissue culture of various tissues of watermelon andregeneration of plants therefrom is well known and widely published. Forexample, reference may be had to Sultana and Rahman, Tissue CultureMethods of Watermelon, (2012); Teng, et al., HortScience, 27:9,1030-1032 (1992); Teng, et al., HortScience, 28:6, 669-1671 (1993);Zhang, et al., Journal of Genetics and Breeding, 46:3, 287-290 (1992);Webb, et al., Plant Cell Tissue and Organ Culture, 38:1, 77-79 (1994);Curtis, et al., Journal of Experimental Botany, 45:279, 1441-1449(1994); Nagata, et al., Journal for the American Society forHorticultural Science, 125:6, 669-672 (2000); and Ibrahim, et al., PlantCell Tissue and Organ Culture, 28(2), 139-145 (1992). It is clear fromthe literature that the state of the art is such that these methods ofobtaining plants are routinely used and have a very high rate ofsuccess. Thus, another aspect of this invention is to provide cellswhich upon growth and differentiation produce watermelon plants havingthe physiological and morphological characteristics of variety SWD8732.

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, meristematic cells, andplant cells that can generate tissue culture that are intact in plantsor parts of plants, such as leaves, pollen, embryos, roots, root tips,anthers, pistils, flowers, seeds, petioles, suckers, and the like. Meansfor preparing and maintaining plant tissue culture are well known in theart. By way of example, a tissue culture comprising organs has been usedto produce regenerated plants. U.S. Pat. Nos. 5,959,185, 5,973,234, and5,977,445 describe certain techniques, the disclosures of which areincorporated herein by reference.

Additional Breeding Methods

This invention also is directed to methods for producing a watermelonplant by crossing a first parent watermelon plant with a second parentwatermelon plant wherein the first or second parent watermelon plant isa watermelon plant of watermelon line CA9. Further, both first andsecond parent watermelon plants can come from watermelon line CA9. Thus,any such methods using watermelon line CA9 are part of this invention:selfing, backcrosses, hybrid production, crosses to populations, and thelike. All plants produced using watermelon line CA9 as at least oneparent are within the scope of this invention, including those developedfrom cultivars derived from watermelon line CA9. Advantageously, thiswatermelon cultivar could be used in crosses with other, different,watermelon plants to produce the first generation (F₁) watermelon hybridseeds and plants with superior characteristics. The cultivar of theinvention can also be used for transformation where exogenous genes areintroduced and expressed by the cultivar of the invention. Geneticvariants created either through traditional breeding methods usingwatermelon line CA9 or through transformation of watermelon line CA9 byany of a number of protocols known to those of skill in the art areintended to be within the scope of this invention.

The following describes breeding methods that may be used withwatermelon line CA9 in the development of further watermelon plants. Onesuch embodiment is a method for developing watermelon line CA9 progenywatermelon plants in a watermelon plant breeding program comprising:obtaining the watermelon plant, or a part thereof, of watermelon lineCA9, utilizing said plant or plant part as a source of breedingmaterial, and selecting a watermelon line CA9 progeny plant withmolecular markers in common with watermelon line CA9 and/or withmorphological and/or physiological characteristics selected from thecharacteristics listed in Table 1. Breeding steps that may be used inthe watermelon plant breeding program include pedigree breeding,backcrossing, mutation breeding, and recurrent selection. In conjunctionwith these steps, techniques such as RFLP-enhanced selection, geneticmarker enhanced selection (for example, SSR markers), and the making ofdouble haploids may be utilized.

Another method involves producing a population of watermelon line CA9progeny watermelon plants, comprising crossing watermelon line CA9 withanother watermelon plant, thereby producing a population of watermelonplants, which, on average, derive 50% of their alleles from watermelonline CA9. A plant of this population may be selected and repeatedlyselfed or sibbed with a watermelon cultivar resulting from thesesuccessive filial generations. One embodiment of this invention is thewatermelon cultivar produced by this method and that has obtained atleast 50% of its alleles from watermelon line CA9.

One of ordinary skill in the art of plant breeding would know how toevaluate the traits of two plant varieties to determine if there is nosignificant difference between the two traits expressed by thosevarieties. For example, see Fehr and Walt, Principles of CultivarDevelopment, pp. 261-286 (1987). Thus the invention includes watermeloncultivar SWD8732 progeny watermelon plants comprising a combination ofat least two watermelon line CA9 traits selected from the groupconsisting of those listed in Table 1, so that said progeny watermelonplant is not significantly different for said traits than watermelonline CA9 as determined at the 5% significance level when grown in thesame environmental conditions. Using techniques described herein,molecular markers may be used to identify said progeny plant as awatermelon line CA9 progeny plant. Mean trait values may be used todetermine whether trait differences are significant, and preferably thetraits are measured on plants grown under the same environmentalconditions. Once such a variety is developed, its value is substantialsince it is important to advance the germplasm base as a whole in orderto maintain or improve traits such as yield, disease resistance, pestresistance, and plant performance in extreme environmental conditions.

Progeny of watermelon line CA9 may also be characterized through theirfilial relationship with watermelon line CA9, as for example, beingwithin a certain number of breeding crosses of watermelon line CA9. Abreeding cross is a cross made to introduce new genetics into theprogeny, and is distinguished from a cross, such as a self or a sibcross, made to select among existing genetic alleles. The lower thenumber of breeding crosses in the pedigree, the closer the relationshipbetween watermelon line CA9 and its progeny. For example, progenyproduced by the methods described herein may be within 1, 2, 3, 4, or 5breeding crosses of watermelon line CA9.

As used herein, the term “plant” includes plant cells, plantprotoplasts, plant cell tissue cultures from which watermelon plants canbe regenerated, plant calli, plant clumps, and plant cells that areintact in plants or parts of plants, such as leaves, pollen, embryos,cotyledons, hypocotyl, roots, root tips, anthers, pistils, flowers,ovules, seeds, stems, and the like.

Tables

Tables 2 and 3 show comparisons of phenotypic characteristics ofwatermelon line CA9 versus a number of commercial and experimentaldiploid watermelon varieties. The phenotypic data trial presented inTables 2 and 3 was conducted at the Sakata Research Station in Woodland,Calif. Watermelon seed was sown on Jul. 1, 2014 and transplanted to thefield on July 16, 2014. Data was collected between Jul. 18, 2014 andSep. 29, 2014. Diploid comparison varieties include experimentalvarieties Koufuku-1-1-1-1-1-1-1-1-1-m, SWD 8732, FWD 8704, FWD 8722, andFWD 8718, and commercial varieties Ace, SP-1, SP-4, Sidekick,Accomplice, Minipool and Mickylee. Watermelon SWD 8732 is a diploidhybrid produced using watermelon line CA9 as a parent.

TABLE 2 Koufuku-1- 1-1-1-1-1-1- VARIETY CA9 1-1-m SWD 8732 FWD 8704 FWD8722 FWD 8718 Ace General Fruit type Oblong Oblong Oblong Oblong OblongOblong Oblong gray Adaptation Most U.S. Most U.S. Most U.S. Most U.S.Most U.S. Most U.S. Most U.S. Areas Areas Areas Areas Areas Areas AreasRelative maturity 70 days 70 days 70 days 70 days 70 days 70 days 70days Ploidy Diploid Diploid Diploid Diploid Diploid Diploid DiploidNumber of 4 including 3 including 3 including 4 including 4 including 3including 4 including main stems at stem holding stem holding stemholding stem holding stem holding stem holding stem holding crown fruitfruit fruit fruit fruit fruit fruit Staminate  8  6  6  5  6  5  8flowers/plant at first fruit set Stem shape Round Round Round RoundRound Round Round Stem diameter 5.0 mm 4.0 mm 6.0 mm 5.0 mm 5.0 mm 5.0mm 6.0 mm at 2nd node Stem surface Pubescent Pubescent PubescentPubescent Pubescent Pubescent Pubescent Leaf shape Ovate Ovate OvateOvate Ovate Ovate Ovate Leaf lobes Lobed Lobed Lobed Lobed Lobed LobedLobed Leaf length 15.0 cm 16.5 cm. 18.0 cm 16.0 cm. 17.0 cm 15.0 cm 18.0cm Leaf width 13.0 cm 14.0 cm 16.0 cm 15.0 cm 14.0 cm 12.0 cm 15.0 cmLeaf size Longer Longer than Longer than Longer than Longer than Longerthan Longer than than wide wide wide wide wide wide wide Dorsal surfacePubescent Pubescent Pubescent Pubescent Pubescent Pubescent Pubescentpubescence Ventral surface Pubescent Pubescent Pubescent PubescentPubescent Pubescent Pubescent pubescence Leaf color RHS 137B RHS 137BRHS 137A RHS 137A RHS 147A RHS 137A RHS 147A (upper); RHS (upper); RHS(upper); RHS (upper); RHS (upper); RHS (upper); RHS (upper); RHS 137C(lower) 137C (lower) 137B (lower) 137B (lower) 147B (lower) 137B (lower)147B (lower) Flower diameter 3.5 cm 3.5 cm 3.2 cm 4.0 cm 3.5 cm 3.2 cm3.5 cm across staminate Flower color RHS 4A RHS 4A RHS 4A RHS 4A RHS 4ARHS 4A RHS 1C with slight with slight with slight with slight withslight with slight with slight RHS 144A RHS 144A RHS 144A RHS 144A RHS144A RHS 144A RHS 144B Mature fruit shape Oval Oval Oval Oval Oval OvalOval Fruit length 14.0 cm 12.0 cm 15.0 cm 17.0 cm 17.0 cm 18.0 cm 17.0cm Fruit diameter 12.0 cm 13.0 cm 14.0 cm 15.0 cm 16.0 cm 15.0 cm 14.0cm at midsection Avg. fruit weight 823 g 806 g 950 g 1084 g 1176 g 1492g 1768 g Max. fruit weight 1167 g 996 g 1089 g 1427 g 1804 g 1730 g 2092g Fruit surface Smooth Smooth Smooth Smooth Smooth Smooth Smooth Skincolor pattern Stripe Stripe Stripe Stripe Stripe Stripe Stripe Primarycolor RHS 146D RHS 146C RHS 146B RHS 146C RHS 146B RHS 146D RHS 146DSecondary color RHS 139A RHS 137A RHS 139A RHS 146A RHS 139A RHS 146BRHS 146A Rind texture Brittle Brittle Brittle Brittle Brittle BrittleBrittle Penetrometer reading 0.96 lb 0.80 lb 0.74 lb 1.2 lb 0.80 lb 0.98lb 1.0 lb Rind thickness 1.0 mm 5.0 mm 1.0 mm 2.0 mm 2.0 mm 2.0 mm 5.0mm blossom end Rind thickness sides 2.0 mm 2.0 mm 2.0 mm 2.0 mm 1.0 mm2.0 mm 2.0 mm Flesh texture Soft Soft Soft Soft Soft Soft Soft Fleshcourseness Course Course Course Course Fine-little Course Course fibrousfibrous fibrous fibrous fiber fibrous fibrous Flesh color RHS 6A RHS179B RHS 15B RHS 179A RHS 18A RHS 179B RHS 179B (Yellow) (Greyed-(Yellow- (Greyed- (Yellow- (Greyed- (Greyed- Red) Orange) Red) Orange)Red) Red) Seed size Medium Medium Medium Medium Small Medium Large Seedlength 9.0 mm 8.0 mm 9.0 mm 9.0 mm 8.0 mm 9.0 mm 11.0 mm Seed width 6.0mm 5.0 mm 6.0 mm 7.0 mm 5.0 mm 5.0 mm 7.0 mm Seed thickness 2.0 mm 2.0mm 2.0 mm 2.0 mm 1.0 mm 2.0 mm 2.0 mm Number of 80 60 60 80 120 80 100seeds per fruit Seed color RHS 200A RHS 166A RHS 200B RHS 200B RHS 202ARHS 200A RHS 200B with 202A at top Known disease Untested UntestedUntested Untested Untested Untested None resistance claimed

TABLE 3 VARIETY CA9 SP-1 SP-4 Sidekick Accomplice Minipool MickyleeGeneral Fruit type Oblong Round gray Round gray Crimson sweet, Oblong,Round gray, Round gray, very small light green large large AdaptationMost U.S. Most U.S. Most U.S. Most U.S. Most U.S. Most U.S. Most U.S.Areas Areas Areas Areas Areas Areas Areas Relative 70 days 75 days 75days 85 days 85 days 85 days 90 days maturity Ploidy Diploid DiploidDiploid Diploid Diploid Diploid Diploid Number of 4 including 3including 3 including 3 including 3 including 4 including 4 includingmain stems at stem holding stem holding stem holding stem holding stemholding stem holding stem holding crown fruit fruit fruit fruit fruitfruit fruit Staminate  8  6  5  6  6  4  5 flowers/plant at first fruitset Stem shape Round Round Round Round Round Round Round Stem diameter5.0 mm 5.0 mm 5.0 mm 4.0 mm 4.0 mm 7.0 mm 5.0 mm at 2nd node Stemsurface Pubescent Pubescent Pubescent Pubescent Pubescent PubescentPubescent Leaf shape Ovate Ovate Ovate Ovate Ovate Ovate Ovate Leaflobes Lobed Lobed Lobed Lobed Lobed Lobed Lobed Leaf length 15.0 cm 20.0cm 17.0 cm 16.0 cm 15.0 cm 17.0 cm 19.0 cm Leaf width 13.0 cm 17.0 cm15.0 cm 12.0 cm 14.0 cm 9.0 cm 12.0 cm Leaf size Longer than Longer thanLonger than Longer than Longer than Longer than Longer than wide widewide wide wide wide wide Dorsal surface Pubescent Pubescent PubescentPubescent Pubescent Pubescent Pubescent pubescence Ventral surfacePubescent Pubescent Pubescent Pubescent Pubescent Pubescent Pubescentpubescence Leaf color RHS 137B RHS 147A RHS 137A RHS 137A RHS 138A RHS137A RHS 137A (upper); RHS (upper); RHS (upper); RHS (upper); RHS(upper); RHS (upper); RHS (upper); RHS 137C (lower) 147B (lower) 137B(lower) 137B (lower) 138B (lower) 137B (lower) 137B (lower) Flowerdiameter 3.5 cm 3.8 cm 2.8 cm 2.5 cm 2.5 cm 3.2 cm 3.5 cm acrossstaminate Flower color RHS 4A RHS 1C RHS 4A RHS 4A RHS 4A RHS 4A RHS 1Cwith slight and with slight with slight with slight with slight and RHS144A RHS 144B RHS 144C RHS 144C RHS 144C RHS 144C RHS 144B Mature fruitshape Oval Round Round Oval Oval Round Round Fruit length 14.0 cm 18.0cm 17.0 cm 10.0 cm 10.0 cm 19.0 cm 20.0 cm Fruit diameter 12.0 cm 17.0cm 14.0 cm 10.0 cm 9.0 cm 17.0 cm 18.0 cm at midsection Avg. fruitweight 823 g 2143 g 1557 g 415 g 415 g 2464 g 2342 g Max. fruit weight1167 g 2174 g 1636 g 426 g 639 g 2653 g 2794 g Fruit surface SmoothSmooth Smooth Smooth Smooth Smooth Smooth Skin color pattern StripeSmall stripe/ Small stripe/ Stripe Solid Small stripe/ Small stripe/mottle/net mottle/net mottle/net mottle/net Primary color RHS 146D RHS146C RHS 147D RHS 137A RHS 147D RHS 145B RHS 145D Secondary color RHS139A RHS 139A RHS 146A RHS 146D RHS 144B RHS 144A RHS 144A Rind textureBrittle Brittle Brittle Brittle Brittle Brittle Brittle Penetrometerreading 0.96 lb 1.4 Lb. 1.2 Lb. 1.8 Lb. 1.6 Lb. 2.0 Lb. 2.5 Lb. Rindthickness 1.0 mm 6.0 mm 1.0 mm 2.0 mm 3.0 mm 5.0 mm 8.0 mm blossom endRind thickness sides 2.0 mm 2.0 mm 1.0 mm 3.0 mm 1.0 mm 10.0 mm 10.0 mmFlesh texture Soft Soft Soft Crisp Crisp Crisp Crisp Flesh coursenessCourse Course Course Fine-little Fine-little Fine-little Fine-littlefibrous fibrous fibrous fiber fiber fiber fiber Flesh color RHS 6A RHS150D RHS 150D RHS 179C RHS 179C RHS 50C RHS 42B (Yellow) (Yellow-(Yellow- (Greyed- (Greyed- (Red) (Red) Green) Green) Red) Red) Seed sizeMedium Large Small Medium Medium Small Medium Seed length 9.0 mm 10.0 mm6.0 mm 10.0 mm 10.0 mm 8.0 mm 8.0 mm Seed width 6.0 mm 6.0 mm 3.0 mm 7.0mm 6.0 mm 5.0 mm 5.0 mm Seed thickness 2.0 mm 2.0 mm 1.0 mm 2.0 mm 2.0mm 2.0 mm 1.0 mm Number of 80 80 100 40 40 120 80 seeds per fruit Seedcolor RHS 200A RHS 200A RHS 200A RHS N167A RHS N167A RHS 202A at RHS200A and and edge and N199D RHS 202A RHS 202A at center Known diseaseUntested Co race 1 and Co race 1, Co race 1 Co race 1, None Fon race 0resistance Fon race 1, 2 Fon race 1, 2 Fon race 0, 1 claimed

Table 4 shows a comparison of the male flower count of watermelon lineCA9 versus a number of commercial and experimental diploid watermelonvarieties. Table 4 shows the date on the first column and the flowercount for the watermelon varieties as indicated. The data presented inTable 4 was obtained in a male flower count trial conducted at theSakata Research Station in Woodland, Calif. Watermelon seed was sown onJuly 1, 2014 and transplanted to the filed on July 16, 2014. Data wascollected between July 18, 2014 and September 29, 2014. Two plants ofeach variety were included in the trial. Comparison varieties includeexperimental varieties Koufuku-1-1-1-1-1-1-1-1-1-m, SWD 8732, FWD 8704,FWD 8722, and FWD 8718, and commercial varieties Ace, SP-1, SP-4,Sidekick, Accomplice, Minipool and Mickylee. Watermelon SWD 8732 is adiploid hybrid produced using watermelon line CA9 as a parent.

TABLE 4 Koufuku- 1-1-1-1-1- SWD FWD FWD FWD VARIETY CA9 1-1-1-1-m 87328704 8722 8718 Ace SP-1 SP-4 Sidekick Accomplice Minipool Mickylee 1-Aug1 2-Aug 2 2 1 3-Aug 1 1 3 1 2 4-Aug 2 2 1 2 1 1 3 5-Aug 2 2 1 1 1 6-Aug3 1 3 2 1 2 1 1 2 7-Aug 1 2 1 1 2 2 4 2 2 1 8-Aug 2 3 1 1 1 1 3 1 1 3 39-Aug 4 2 2 2 2 2 2 3 3 1 10-Aug 1 2 1 3 2 2 3 1 4 1 3 2 11-Aug 1 4 2 32 3 4 6 3 1 6 2 12-Aug 3 1 1 7 3 1 4 3 5 3 3 7 0 13-Aug 6 4 1 4 2 4 7 55 3 4 4 4 14-Aug 5 5 1 6 5 6 6 6 9 5 4 8 6 15-Aug 3 7 2 4 4 5 7 7 8 10 67 3 16-Aug 7 7 1 13 3 3 9 8 6 3 7 6 5 17-Aug 6 6 2 9 8 6 9 8 10 7 1 7 518-Aug 7 6 3 10 9 3 10 11 9 10 6 8 7 19-Aug 0 5 2 17 11 7 14 15 4 15 197 6 20-Aug 2 10 4 11 8 6 10 8 18 15 25 12 6 21-Aug 5 10 7 10 6 9 9 16 1023 16 11 12 22-Aug 7 13 4 19 8 7 13 19 20 22 21 9 8 23-Aug 6 13 6 18 1213 14 38 33 30 37 10 11 24-Aug 21 26 9 22 16 14 27 51 38 42 46 16 1825-Aug 16 21 14 34 19 23 29 77 54 47 51 17 10 26-Aug 35 30 16 38 28 2241 87 53 58 64 29 14 27-Aug 25 22 20 23 20 18 33 79 52 69 59 32 9 28-Aug37 19 33 24 21 15 27 76 61 65 58 29 13 29-Aug 31 25 54 28 31 30 34 89 7089 85 41 21 30-Aug 59 28 37 30 35 29 27 102 73 62 63 52 27 31-Aug 62 3737 36 41 34 29 118 80 70 82 53 20 1-Sep 73 22 41 42 38 24 30 124 66 5578 58 27 2-Sep 81 29 37 34 32 32 22 111 78 81 96 41 31 3-Sep 82 25 44 4724 29 22 122 83 66 107 64 33 4-Sep 82 22 44 39 31 32 17 132 98 93 115 5939 5-Sep 70 17 46 37 27 25 9 108 76 51 87 46 37 6-Sep 88 19 47 41 29 3112 112 89 50 83 58 47 7-Sep 83 20 55 34 33 30 17 101 76 55 78 42 518-Sep 97 20 50 36 34 40 15 79 70 53 61 35 50 9-Sep 103 19 55 32 40 28 1076 45 32 46 24 47 10-Sep 89 14 48 31 36 30 10 50 47 36 44 30 40 11-Sep63 12 41 20 23 28 14 37 27 20 44 17 39 12-Sep 50 8 35 21 22 31 11 41 2417 26 6 31 13-Sep 56 10 30 18 31 34 12 46 18 13 14 23 27 14-Sep 64 12 3023 30 25 17 32 12 12 27 11 35 15-Sep 46 9 31 16 24 24 16 18 11 10 10 933 16-Sep 17-Sep 29 7 23 14 21 22 14 13 10 5 3 5 20 18-Sep 16 6 12 11 1316 6 9 9 1 0 0 17 19-Sep 9 6 12 7 14 14 10 8 6 1 1 2 15 20-Sep 13 12 1220 21 22 8 17 4 2 0 1 14 21-Sep 8 9 13 8 10 13 4 9 6 0 0 3 12 22-Sep 9 77 15 16 12 7 8 4 0 0 4 16 23-Sep 8 4 5 11 14 14 5 5 3 4 0 3 7 TOTAL (21566 625 978 937 866 826 667 2094 1496 1310 1587 932 879 Plants) TOTAL(1 783 312.5 489 468.5 433 413 333.5 1047 748 655 793.5 466 439.5 Plant)MEAN 15.06 6.01 9.40 9.01 8.33 7.94 6.41 20.1 14.3 12.59 15.26 8.96 8.45

As shown in Table 4, watermelon line CA9 produces the third highestflower count when compared to commercial and experimental diploidwatermelon varieties.

Watermelon line CA9 was used as a parent in a cross to produce a diploidhybrid watermelon pollenizer designated SWD 8732. Table 5 shows acomparison of male flower ratings of SWD 8732 versus experimental andcommercial diploid pollenizer lines. Table 5 shows the date on the firstcolumn and the male flower rating for the watermelon varieties asindicated. The data presented in Table 5 was obtained in a male flowercount trial conducted at the Sakata Research Station in Ft. Myers,Florida in spring of 2009. The trial was transplanted on March 18, 2009and male flowering ratings were taken daily from April 8, 2009 until May11, 2009. Comparison varieties include experimental variety FWD 8700 andcommercial varieties Ace, SP-1, Sidekick, Minipool, SP-4 and Companion.Male flowering was rated through visual observation using a 0-5 ratingscale in which 5 is very high male flower quantity, 4 is high maleflower quantity, 3 is average male flower quantity, 2 is low male flowerquantity and 1 is very low male flower quantity. For the data in Table5, each plot had 20 plants.

TABLE 5 VARIETY SWD 8732 FWD 8700 Ace SP-1 Sidekick Minipool SP-4Companion 8-Apr 2.5 3 3.5 2.5 0 0 2 0 9-Apr 2.5 4.5 3 1.5 1 0 1.5 010-Apr 3 3.5 3 1.5 1 0.5 1.5 1.5 11-Apr 3.5 3 4.5 3.5 3 3 4 2.5 12-Apr4.5 5.5 4.5 4 3 2 3 2.5 13-Apr 4.5 5.5 4.5 4 2.5 2.5 4 3 14-Apr 4.5 54.5 3.5 2.5 2.5 3.5 3 15-Apr 3.5 4.5 4.5 3.5 2.5 3 3.5 2.5 16-Apr 3.54.5 4 3.5 2.5 3 3.5 3 17-Apr 3.5 4 4 4 3 3.5 3.5 2.5 18-Apr 4.5 4 4.53.5 3 3.5 3 2.5 19-Apr 4.5 4.5 4 3.5 4 4 3.5 2.5 20-Apr 4.5 4 4 4 3.53.5 4 3.5 21-Apr 4.5 4 4.5 4 3.5 3.5 3.5 3 22-Apr 3.5 4 3.5 4 3.5 3.53.5 3 23-Apr 3.5 4.5 4 4 4 4 3.5 3.5 24-Apr 3.5 4 3.5 4 4 3.5 3.5 325-Apr 3 4 3.5 4 4 3.5 3.5 3 26-Apr 3 4 3.5 4 4 4 3.5 3.5 27-Apr 4 4 44.5 4 4 3.5 4 28-Apr 4 4 4 4 4 4 3 3.5 29-Apr 4 3.5 4 4 3.5 3.5 3.5 3.530-Apr 3 3 2.5 4 3.5 2.5 2 3 1-May 3 3 2.5 3.5 3.5 3 2.5 3 2-May 3.5 3 23 3 3 2 2.5 3-May 3 3 2.5 3 3 2.5 2 2 4-May 3 3.5 1 3 2.5 3 2 2 5-May 33.5 1.5 3 3.5 2.5 2 2 6-May 3.5 3.5 1.5 2 2.5 1.5 1 1.5 7-May 3.5 3 2.52 3.5 2.5 2 2 8-May 3.5 3 1.5 2.5 2.5 3 1.5 1.5 9-May 2.5 2 1.5 2.5 2.52.5 1.5 1.5 10-May 2.5 2.5 2.5 2 2.5 2.5 1.5 1 11-May 2 3 2 2 2.5 2.51.5 1 Average 3.5 4 3.3 3.3 3 2.8 2.7 2.4

Watermelon line CA9 was used as a parent in a cross to produce a diploidhybrid watermelon pollenizer designated SWD 8732. Table 6 shows acomparison of seedless triploid watermelon fruit produced using BoldRuler as the field variety and either diploid watermelon SWD 8732 or Aceas the pollenizer plant. Two blocks totaling a quarter acre were plantedfor each experiment. The trial was conducted at the Sakata ResearchStation in Ft. Myers, Florida in spring of 2013.

TABLE 6 Triploid fruit Diploid pollenizer produced per acre DifferenceSWD 8732 3948 +384 Ace 3564

As shown in Table 6, Bold Ruler produces 384 more fruit per acre whenpollenized with SWD 8732 versus when pollenized with Ace. In addition,the fruit size of SWD 8732 seemed a little smaller than Ace on averageat the time of evaluation and the vine of SWD 8732 was larger and morevigorous and helped to provide better coverage of Bold Ruler fruit.

Watermelon SWD 8732 is a diploid hybrid produced using watermelon lineCA9 as a parent. Table 7 shows a comparison of the total number ofmarketable seedless triploid watermelon fruits produced using SWD 8732as the diploid pollenizer plant versus the fruit produced usingexperimental or commercial diploid varieties as pollenizers of fieldvariety Sweet Treasure. The trial was conducted at the Sakata ResearchStation in Ft. Myers, Florida in fall of 2010. The trial plots each hadfour row beds and each row had 15 Sweet Treasure (triploid) plants. Onepollenizer plant was interplanted after every five triploid plants. Intotal, 60 Sweet Treasure plants and 9 pollenizer plants were used foreach plot. Between two plots, five foot border plot spaces were insertedand only Sweet Treasure without pollenizer were transplanted; fruitsfrom this space were not counted. Comparison varieties includeexperimental varieties FWD 8700, FWD 8709, FWD 8704 and FWD 8710 andcommercial varieties Sweet Harmony, Valentino, Ace, SP-5 and Sidekick.Table 7, column 1 shows the variety used as the diploid pollenizer,column 2 shows the total number of fruits produced, column 3 shows thetotal number of marketable seedless triploid watermelon fruits produced,column 4 shows the total number of culls, and column 5 shows the totalnumber of diploid pollenizer fruits produced.

TABLE 7 Marketable # Total # of Total # of triploid Total # diploidpollenizer Pollenizer of fruits fruits of culls fruits SWD 8732 151 9257 55 FWD 8700 110 64 44 44 FWD 8709 138 67 68 54 FWD 8704 124 66 58 42FWD 8710 133 48 80 30 Sweet Harmony 139 90 49 20 Valentino 142 53 82 25Ace 153 49 101 33 SP-5 145 63 80 25 Sidekick 141 59 82 85

The use of the terms “a,” “an,” and “the,” and similar referents in thecontext of describing the invention (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext. The terms “comprising,” “having,” “including,” and “containing”are to be construed as open-ended terms (i.e., meaning “including, butnot limited to,”) unless otherwise noted. Recitation of ranges of valuesherein are merely intended to serve as a shorthand method of referringindividually to each separate value falling within the range, unlessotherwise indicated herein, and each separate value is incorporated intothe specification as if it were individually recited herein. Forexample, if the range 10-15 is disclosed, then 11, 12, 13, and 14 arealso disclosed. All methods described herein can be performed in anysuitable order unless otherwise indicated herein or otherwise clearlycontradicted by context. The use of any and all examples, or exemplarylanguage (e.g., “such as”) provided herein, is intended merely to betterilluminate the invention and does not pose a limitation on the scope ofthe invention unless otherwise claimed. No language in the specificationshould be construed as indicating any non-claimed element as essentialto the practice of the invention.

Deposit Information

A deposit of the watermelon seed of this invention is maintained bySakata Seed America, Inc., 18095 Serene Drive, Morgan Hill, Calif.95037, U.S.A. Access to this deposit will be available during thependency of this application to persons determined by the Commissionerof Patents and Trademarks to be entitled thereto under 37 CFR §1.14 and35 USC §122. Upon allowance of any claims in this application, allrestrictions on the availability to the public of the variety will beirrevocably removed by affording access to a deposit of at least 2,500seeds of the same variety with the American Type Culture Collection(ATCC), 10801 University Boulevard, Manassas, Va. 20110 or NationalCollections of Industrial, Food and Marine Bacteria (NCIMB), 23 StMachar Drive, Aberdeen, Scotland, AB24 3RY, United Kingdom.

While a number of exemplary aspects and embodiments have been discussedabove, those of skill in the art will recognize certain modifications,permutations, additions, and sub-combinations thereof. It is thereforeintended that the following appended claims and claims hereafterintroduced are interpreted to include all such modifications,permutations, additions, and sub-combinations as are within their truespirit and scope.

1. A seed of watermelon line CA9, wherein a representative sample ofseed of said line was deposited under ATCC Accession No. PTA ______. 2.A watermelon plant, or a part thereof, produced by growing the seed ofclaim
 1. 3. A watermelon plant having all the physiological andmorphological characteristics of the watermelon plant of claim
 2. 4. Atissue culture produced from protoplasts or cells from the plant ofclaim 2, wherein said cells or protoplasts are produced from a plantpart selected from the group consisting of leaf, pollen, embryo,cotyledon, hypocotyl, meristematic cell root, root tip, pistil, anther,ovule, flower, shoot, stem, seed, and petiole wherein said seed is notan F₁ seed.
 5. A watermelon plant regenerated from the tissue culture ofclaim 4, wherein the plant has all of the morphological andphysiological characteristics of watermelon line CA9.
 6. A method forproducing a watermelon seed, said method comprising crossing twowatermelon plants and harvesting the resultant watermelon seed, whereinat least one watermelon plant is the watermelon plant of claim
 2. 7. AnF₁ watermelon seed produced by the method of claim
 6. 8. An F iwatermelon plant produced by growing said seed of claim
 7. 9. (canceled)10. The method of claim 6, wherein one of said watermelon plants iswatermelon line CA9 and the other is transgenic.
 11. A method ofproducing an herbicide resistant watermelon plant, wherein said methodcomprises introducing a gene conferring herbicide resistance into theplant of claim 2, wherein the gene is selected from the group consistingof glyphosate, sulfonylurea, imidazolinone, dicamba, glufosinate,phenoxy proprionic acid, L-phosphinothricin, cyclohexone,cyclohexanedione, triazinc, and benzonitrile.
 12. An herbicide resistantwatermelon plant produced by the method of claim 11, wherein said plantotherwise has all of the physiological and morphological characteristicsof watermelon line CA9.
 13. A method of producing a pest of insectresistant watermelon plant, wherein said method comprises introducing agene conferring pest or insect resistance into the plant of claim
 2. 14.A pest or insect resistant watermelon plant produced by the method ofclaim 13, wherein said plant otherwise has all of the physiological andmorphological characteristics of watermelon line CA9.
 15. The watermelonplant of claim 14, wherein the gene encodes a Bacillus thuringiensisendotoxin, and wherein said plant otherwise has all of the physiologicaland morphological characteristics of watermelon line CA9.
 16. A methodof producing a disease resistant watermelon plant, wherein said methodcomprises introducing a gene conferring disease resistance into theplant of claim
 2. 17. A disease resistant watermelon plant produced bythe method of claim 16, wherein said plant otherwise has all of thephysiological and morphological characteristics of watermelon line CA9.18. A method of producing a watermelon plant with a value-added trait,wherein said method comprises introducing a gene conferring avalue-added trait into the plant of claim 2, and wherein said geneencodes a protein selected from the group consisting of a ferritin, anitrate reductase, a monellin, fructosyltransferase, levansucrase,α-amylase, invertasc and starch branching enzyme or encoding anantisense of stearyl-ACP desaturase.
 19. A watermelon plant with avalue-added trait produced by the method of claim 18, wherein said plantotherwise has all of the physiological and morphological characteristicsof watermelon line CA9.
 20. A method of introducing a desired trait intowatermelon line CA9, wherein the method comprises: (a) crossing a plantof watermelon line CA9, wherein a representative sample of seed wasdeposited under ATCC Accession No. PTA-______, with a plant of anotherwatermelon cultivar that comprises a desired trait to produce progenyplants, and wherein the desired trait is selected from the groupconsisting of male sterility, herbicide resistance, insect or pestresistance, modified bolting and resistance to bacterial disease, fungaldisease or viral disease, (b) selecting one of more progeny plants thathave the desired trait; (c) backcrossing the selected progeny plantswith watermelon line CA9 to produce backcross progeny plants; (d)selecting for backcross progeny plants that have the desired trait; and(e) repeating steps (c) and (d) two or more times in succession toproduce selected third or higliei backcross progeny plants that comprisethe desired trait.
 21. A watermelon plant produced by the method ofclaim 70, wherein the plant has the desired trait and otherwise all ofthe physiological and morphological characteristics of watermelon lineCA9.
 22. The watermelon plant of claim 21, wherein the desired trait isherbicide resistance and the resistance is conferred to an herbicideselected from the group consisting of glyphosate, sulfonylurea,imidazolinone, dicamba, glufosinatc, phenoxy proprionic acid,L-phosphinothricin, cyclohexone, cyclohexanedione, triazine, andbenzonitrile.
 23. The watermelon plant of claim 21, wherein the desiredtrait is insect or pest resistance and the insect or pest resistance isconferred by a transgene encoding a Bacillus thuringiensis endotoxin.24. A method for producing triploid, seedless watermelon fruit, whereinthe method comprises the steps of: (a) Planting a field with triploidwatermelon plants; (b) Obtaining diploid pollenizer watermelon plantsfor pollinizing triploid watermelon plants, wherein at least one parentof said diploid pollenizer watermelon plants is the plant of claim 2;(c) Planting said pollenizer watermelon plants in the field of triploidwatermelon plants; (d) Allowing pollination of said triploid watermelonplants by pollen of said pollenizer watermelon plants to obtaintriploid, seedless watermelon fruit; and (e) Harvesting said triploid,seedless watermelon fruit.
 25. The method for producing triploid,seedless watermelon fruit according to claim 24, whcrein planting ofsaid diploid pollemzer plants is at a ratio of approximately equal to orless than 1 diploid pollenizer watermelon plant to 3 triploid watermelonplants.
 26. The method for producing triploid, seedless watermelon fruitaccording to claim 24, wherein planting of said diploid pollenizerplants is at a ratio of approximately equal to or less than 1 diploidpollenizer watermelon plant to 4 triploid watermelon plants.
 27. Themethod for producing triploid, seedless watermelon fruit according toclaim 24, wherein planting of said diploid pollenizer plants is at aratio of approximately equal to or less than 1 diploid pollenizerwatermelon plant to 5 triploid watermelon plants.
 28. The method forproducing triploid, seedless watermelon fruit according to claim 24,wherein planting of said diploid pollenizer plants is at a ratio ofapproximately equal to or less than 1 diploid pollenizer watermelonplant to 6 triploid watermelon plants
 29. A method for producingtriploid, seedless watermelon fruit, wherein the method comprises thesteps of: (a) Inter-planting pollenizer watermelon plants and triploidwatermelon plants in a field, wherein at least one parent of saidpollenizer watermelon plants is the plant of claim 2; and (b) Allowingpollination of said triploid watermelon plants by pollen of saidpollenizer watermelon plants to obtain triploid, seedless watermelonfruit.
 30. The method for producing triploid, seedless watermelon fruitaccording to claim 29, further comprising harvesting said triploid,seedless watermelon fruit.
 31. A method of producing a graftedwatermelon plant comprising grafting a watermelon scion onto a suitablerootstock, wherein the plant of claim 2 is used as the scion.